Advertisement

Intranasal immunization with coxsackievirus A16 virus-like particles confers protection against lethal infection in neonatal mice

  • Xiangpeng Chen
  • Yong Zhang
  • Naiying Mao
  • Shuangli Zhu
  • Tianjiao Ji
  • Wenbo XuEmail author
Original Article

Abstract

Coxsackievirus A16 (CV-A16) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children and has become prevalent in the Asia-Pacific region in recent years. However, no approved vaccines or drugs are available for CV-A16 infection. CV-A16 virus-like particles (VLPs) are a potential vaccine candidate; however, whether the intranasal route of immunization is suitable for inducing immune responses against CV-A16 infection has not been clarified. In this study, the comprehensive immunogenicity and protective efficacy of the CV-A16 VLP vaccine were evaluated by multiple methods in a mouse model. In mice, a high neutralizing antibody (NTAb) titre could be elicited by intranasal immunization with CV-A16 VLPs, which produced NTAb levels similar to those induced by intranasal immunization with inactivated CV-A16. Passive immunity with NTAbs provided very good protection, as the survival rate of the immunized neonatal mice was 100% after challenges with CV-A16 at a dose of 1000 LD50. Passive protective effects were transferred to the neonates via the mother, thus protecting all the pups against challenges with the homologous or heterologous strains of CV-A16 at a dose of 1000 LD50. In addition, intranasal immunization with CV-A16 VLPs also induced the production of mucosal secretory IgA (s-IgA) antibodies, which may inhibit CV-A16 virus invasion. This study provides valuable supplemental information to facilitate our understanding of the specific protective efficacy of CV-A16 VLPs and has significance for development of the candidate vaccine into a safe and effective vaccine.

Notes

Author contributions

XC and WX conceived and designed the study; XC performed the experiments; YZ performed the data analyses; NM performed the ELISPOT analyses; SZ and TJ performed the ELISAs and analysed and interpreted the data. The manuscript was written by XC and proofread by WX. All authors revised the manuscript and approved the final version.

Funding

This work was supported by the Beijing Natural Science Foundation (Grant no. 7184208), the National Science and Technology Major Project of the Ministry of Science and Technology of China (Grant nos. 2018ZX10201002-003-009, 2017ZX10104001-005-010, 2018ZX10713002, and 2018ZX10713001-003), the Basic and Clinical Research Cooperation Project of Capital Medical University (Grant no. 17JL11), and the Research Training Fund of Capital Medical University (Grant no. PYZ2017012). The sponsors played no role in the study design, data analysis, manuscript preparation, or decision to publish.

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Animal and human rights statement

The program for immunization with CV-A16 VLPs or inactivated CV-A16 combined with CpG ODN was approved by the Ethical Committee of the National Institute of Viral Disease Control and Prevention, China CDC. The animal care and use protocol in this study abided by “The Guidance on Treating Experimental Animals”, which was promulgated by the Ministry of Science and Technology of China. No nonhuman primates were used in this study.

References

  1. 1.
    Liu W, Wu S, Xiong Y, Li T, Wen Z, Yan M, Qin K, Liu Y, Wu J (2014) Co-circulation and genomic recombination of coxsackievirus A16 and enterovirus 71 during a large outbreak of hand, foot, and mouth disease in Central China. PLoS One 9:e96051.  https://doi.org/10.1371/journal.pone.0096051 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Liu Y, Wang X, Pang C, Yuan Z, Li H, Xue F (2015) Spatio-temporal analysis of the relationship between climate and hand, foot, and mouth disease in Shandong province, China, 2008–2012. BMC Infect Dis 15:146.  https://doi.org/10.1186/s12879-015-0901-4 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Ang LW, Koh BK, Chan KP, Chua LT, James L, Goh KT (2009) Epidemiology and control of hand, foot and mouth disease in Singapore, 2001–2007. Ann Acad Med Singap 38:106–112PubMedGoogle Scholar
  4. 4.
    Chen JF, Zhang RS, Ou XH, Chen FM, Sun BC (2014) The role of enterovirus 71 and coxsackievirus A strains in a large outbreak of hand, foot, and mouth disease in 2012 in Changsha, China. Int J Infect Dis 28:17–25.  https://doi.org/10.1016/j.ijid.2014.07.024 CrossRefPubMedGoogle Scholar
  5. 5.
    Xing W, Liao Q, Viboud C, Zhang J, Sun J, Wu JT, Chang Z, Liu F, Fang VJ, Zheng Y, Cowling BJ, Varma JK, Farrar JJ, Leung GM, Yu H (2014) Hand, foot, and mouth disease in China, 2008–12: an epidemiological study. Lancet Infect Dis 14:308–318.  https://doi.org/10.1016/S1473-3099(13)70342-6 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Du J, Wang X, Hu Y, Li Z, Li Y, Sun S, Yang F, Jin Q (2014) Changing aetiology of hand, foot and mouth disease in Linyi, China, 2009–2011. Clin Microbiol Infect 20:O47–O49.  https://doi.org/10.1111/1469-0691.12301 CrossRefPubMedGoogle Scholar
  7. 7.
    Sun LM, Wu SL, Tan XH, Li H, Yang F, Zeng HR, Zheng HY, Liu L, He JF (2018) Epidemiological characteristics of Coxsackie virus A16 caused hand foot and mouth disease cases in Guangdong province, 2012–2016. Zhonghua Liu Xing Bing Xue Za Zhi 39:342–346.  https://doi.org/10.3760/cma.j.issn.0254-6450.2018.03.018 CrossRefPubMedGoogle Scholar
  8. 8.
    Yee PTI, Laa Poh C (2017) Impact of genetic changes, pathogenicity and antigenicity on Enterovirus-A71 vaccine development. Virology 506:121–129.  https://doi.org/10.1016/j.virol.2017.03.017 CrossRefPubMedGoogle Scholar
  9. 9.
    Mao Q, Wang Y, Yao X, Bian L, Wu X, Xu M, Liang Z (2014) Coxsackievirus A16: epidemiology, diagnosis, and vaccine. Hum Vaccin Immunother 10:360–367.  https://doi.org/10.4161/hv.27087 CrossRefPubMedGoogle Scholar
  10. 10.
    Wang CY, Li LuF, Wu MH, Lee CY, Huang LM (2004) Fatal coxsackievirus A16 infection. Pediatr Infect Dis J 23:275–276CrossRefGoogle Scholar
  11. 11.
    Chackerian B (2007) Virus-like particles: flexible platforms for vaccine development. Expert Rev Vaccines 6:381–390.  https://doi.org/10.1586/14760584.6.3.381 CrossRefPubMedGoogle Scholar
  12. 12.
    Li HZ, Gang HY, Sun QM, Liu X, Ma YB, Sun MS, Dai CB (2004) Production in Pichia pastoris and characterization of genetic engineered chimeric HBV/HEV virus-like particles. Chin Med Sci J 19:78–83PubMedGoogle Scholar
  13. 13.
    McCormack PL (2014) Quadrivalent human papillomavirus (types 6, 11, 16, 18) recombinant vaccine (gardasil((R))): a review of its use in the prevention of premalignant anogenital lesions, cervical and anal cancers, and genital warts. Drugs 74:1253–1283.  https://doi.org/10.1007/s40265-014-0255-z CrossRefPubMedGoogle Scholar
  14. 14.
    Siddiqui MA, Perry CM (2006) Human papillomavirus quadrivalent (types 6, 11, 16, 18) recombinant vaccine (Gardasil). Drugs 66:1263–1271 (discussion 1272–1263) CrossRefGoogle Scholar
  15. 15.
    Szarewski A (2010) HPV vaccine: cervarix. Expert Opin Biol Ther 10:477–487.  https://doi.org/10.1517/14712591003601944 CrossRefPubMedGoogle Scholar
  16. 16.
    Chung YC, Ho MS, Wu JC, Chen WJ, Huang JH, Chou ST, Hu YC (2008) Immunization with virus-like particles of enterovirus 71 elicits potent immune responses and protects mice against lethal challenge. Vaccine 26:1855–1862CrossRefGoogle Scholar
  17. 17.
    Cao L, Mao F, Pang Z, Yi Y, Qiu F, Tian R, Meng Q, Jia Z, Bi S (2015) Protective effect of enterovirus71 (EV71) viruslike particle vaccine against lethal EV71 infection in a neonatal mouse model. Mol Med Rep 12:2473–2480.  https://doi.org/10.3892/mmr.2015.3680 CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Zhang C, Liu Q, Ku Z, Hu Y, Ye X, Zhang Y, Huang Z (2016) Coxsackievirus A16-like particles produced in Pichia pastoris elicit high-titer neutralizing antibodies and confer protection against lethal viral challenge in mice. Antivir Res 129:47–51.  https://doi.org/10.1016/j.antiviral.2016.02.011 CrossRefPubMedGoogle Scholar
  19. 19.
    Feng Q, He Y, Lu J (2016) Virus-like particles produced in Pichia pastoris induce protective immune responses against coxsackievirus A16 in mice. Med Sci Monit Int Med J Exp Clin Res 22:3370–3382Google Scholar
  20. 20.
    Chen XP, Mao NY, Zhang Y, Xie ZD, Xu WB (2014) Preparation and immunogenicity of virus-like particles of Coxsackievirus A16. Chin J Biol 27:1361–1374Google Scholar
  21. 21.
    Chen XP, Tan XJ, Zhang Y, Xu WB (2014) Immunoprotective effect of inactivated coxsackievirus A16 vaccine in mice. Bing Du Xue Bao 30:226–232PubMedGoogle Scholar
  22. 22.
    Melnick JL, Wimberly IL (1985) Lyophilized combination pools of enterovirus equine antisera: new LBM pools prepared from reserves of antisera stored frozen for two decades. Bull World Health Organ 63:543–550PubMedPubMedCentralGoogle Scholar
  23. 23.
    Reed LJ, Muench H (1938) A simple method of estimating 50% end-points. Am J Hyg 27:493–497Google Scholar
  24. 24.
    Mao Q, Wang Y, Gao R, Shao J, Yao X, Lang S, Wang C, Mao P, Liang Z, Wang J (2012) A neonatal mouse model of coxsackievirus A16 for vaccine evaluation. J Virol 86:11967–11976.  https://doi.org/10.1128/JVI.00902-12 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Cui A, Xu C, Tan X, Zhang Y, Zhu Z, Mao N, Lu Y, Xu W (2013) The development and application of the two real-time RT-PCR assays to detect the pathogen of HFMD. PLoS One 8:e61451.  https://doi.org/10.1371/journal.pone.0061451 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Illum L, Jabbal-Gill I, Hinchcliffe M, Fisher AN, Davis SS (2001) Chitosan as a novel nasal delivery system for vaccines. Adv Drug Deliv Rev 51:81–96CrossRefGoogle Scholar
  27. 27.
    Kemble G, Greenberg H (2003) Novel generations of influenza vaccines. Vaccine 21:1789–1795CrossRefGoogle Scholar
  28. 28.
    Nantachit N, Sunintaboon P, Ubol S (2016) Responses of primary human nasal epithelial cells to EDIII-DENV stimulation: the first step to intranasal dengue vaccination. Virol J 13:142.  https://doi.org/10.1186/s12985-016-0598-z CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Rose MA, Zielen S, Baumann U (2012) Mucosal immunity and nasal influenza vaccination. Expert Rev Vaccines 11:595–607.  https://doi.org/10.1586/erv.12.31 CrossRefPubMedGoogle Scholar
  30. 30.
    Wu HY, Nguyen HH, Russell MW (1997) Nasal lymphoid tissue (NALT) as a mucosal immune inductive site. Scand J Immunol 46:506–513CrossRefGoogle Scholar
  31. 31.
    Kuper CF, Koornstra PJ, Hameleers DM, Biewenga J, Spit BJ, Duijvestijn AM, van Breda Vriesman PJ, Sminia T (1992) The role of nasopharyngeal lymphoid tissue. Immunol Today 13:219–224CrossRefGoogle Scholar
  32. 32.
    Brandtzaeg P, Johansen FE (2005) Mucosal B cells: phenotypic characteristics, transcriptional regulation, and homing properties. Immunol Rev 206:32–63.  https://doi.org/10.1111/j.0105-2896.2005.00283.x CrossRefPubMedGoogle Scholar
  33. 33.
    Kiyono H, Fukuyama S (2004) NALT-versus Peyer’s-patch-mediated mucosal immunity. Nat Rev Immunol 4:699–710.  https://doi.org/10.1038/nri1439 CrossRefPubMedGoogle Scholar
  34. 34.
    Birk R, Aderhold C, Hormann K, Wenzel A, Kramer B, Eschenhagen T, Sommer JU (2016) CpG-oligodeoxynucleotides in chronic rhinosinusitis cell culture. In Vivo 30:47–52PubMedGoogle Scholar
  35. 35.
    Meng Z, Zhang X, Pei R, Zhang E, Kemper T, Vollmer J, Davis HL, Glebe D, Gerlich W, Roggendorf M, Lu M (2016) Combination therapy including CpG oligodeoxynucleotides and entecavir induces early viral response and enhanced inhibition of viral replication in a woodchuck model of chronic hepadnaviral infection. Antivir Res 125:14–24.  https://doi.org/10.1016/j.antiviral.2015.11.001 CrossRefPubMedGoogle Scholar
  36. 36.
    Xiang XX, Zhou XQ, Wang JX, Xie Q, Cai X, Yu H, Zhou HJ (2011) Effects of CpG-ODNs on phenotype and function of monocyte-derived dendritic cells in chronic hepatitis B. World J Gastroenterol 17:4825–4830.  https://doi.org/10.3748/wjg.v17.i43.4825 CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Lin YL, Chow YH, Huang LM, Hsieh SM, Cheng PY, Hu KC, Chiang BL (2018) A CpG-adjuvanted intranasal enterovirus 71 vaccine elicits mucosal and systemic immune responses and protects human SCARB2-transgenic mice against lethal challenge. Sci Rep 8:10713.  https://doi.org/10.1038/s41598-018-28281-5 CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Liu Q, Yan K, Feng Y, Huang X, Ku Z, Cai Y, Liu F, Shi J, Huang Z (2012) A virus-like particle vaccine for coxsackievirus A16 potently elicits neutralizing antibodies that protect mice against lethal challenge. Vaccine 30:6642–6648.  https://doi.org/10.1016/j.vaccine.2012.08.071 CrossRefPubMedGoogle Scholar
  39. 39.
    Emeny RT, Wheeler CM, Jansen KU, Hunt WC, Fu TM, Smith JF, MacMullen S, Esser MT, Paliard X (2002) Priming of human papillomavirus type 11-specific humoral and cellular immune responses in college-aged women with a virus-like particle vaccine. J Virol 76:7832–7842.  https://doi.org/10.1128/jvi.76.15.7832-7842.2002 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Laboratory of Infection and Virology, Beijing Pediatric Research Institute, Beijing Children’s HospitalCapital Medical University, National Center for Children’s HealthBeijingChina
  2. 2.WHO WPRO Regional Polio Reference Laboratory and NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijingChina
  3. 3.WHO WPRO Regional Reference Measles/Rubella Laboratory and NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijingChina

Personalised recommendations